CN117143227B - Specific antibody for combining coronavirus SARS-CoV-2 and SARS-CoV nucleocapsid protein and application thereof - Google Patents
Specific antibody for combining coronavirus SARS-CoV-2 and SARS-CoV nucleocapsid protein and application thereof Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
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- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/08—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses
- C07K16/10—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses from RNA viruses
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Abstract
The invention discloses a specific antibody combined with coronavirus SARS-CoV-2 and SARS-CoV nucleocapsid protein (Nucleocapsid protein) and application thereof. The sequences of CDRs of the antibody are shown in SEQ ID NO.1 to SEQ ID NO. 6. The antibody provided by the invention comprises CDRs with specific sequences, can be specifically combined with SARS-CoV-2 and SARS-CoV nucleocapsid proteins, and has weak combination reaction with nucleocapsid proteins of another highly pathogenic coronavirus MERS-CoV. The antibody provided by the invention has strong targeting property and high binding activity, and the antibody prepared by the preparation method provided by the invention has high purity and can be prepared in a large amount. Overall, the antibodies of the invention can be used for the development and optimization of SARS-CoV and SARS-CoV-2 infection detection methods.
Description
Technical Field
The invention belongs to the technical field of biological medicine, and in particular relates to a specific antibody combined with coronavirus SARS-CoV-2 and SARS-CoV nucleocapsid protein (nucleocap apsi dprotein) and a preparation method and application thereof.
Background
SARS-CoV and SARS-CoV-2 belong to the genus sabbevirus (Sarbecovirus), which are highly pathogenic coronaviruses that infect humans, and mainly encode four structural proteins, spike protein (Spike protein), membrane protein (Membrane protein), envelope protein (Envelope protein) and nucleocapsid protein (nucleocapsid protein). The nucleocapsid protein can bind to viral genomic RNA to form ribonucleocapsid complexes, and can also be involved in viral genome protection, viral RNA replication, and assembly. Nucleocapsid protein (Nucleocapsid protein) is highly conserved and is also the most abundant structural protein expressed on SARS-CoV and SARS-CoV-2. Nucleocapsid proteins (nucleocapsid proteins) are therefore often used in the development of test kits to assess the infection of a patient by the expression level of nucleocapsid protein genes or proteins. Development of antibodies against nucleocapsid proteins (nucleocaps) would facilitate development of methods for detection of SARS-CoV and SARS-CoV-2 infection.
SARS-CoV and SARS-CoV-2 are currently reported to infect human saber virus (Sarbecovirus). Nucleocapsid protein (nucleocap) is one of the structural proteins of coronaviruses, involved in viral replication and ribonucleocapsid formation, and has high conservation, and its gene and viral expression level can be used as an index for detecting viral infection and replication capacity. Antibodies targeting nucleocapsid proteins (nucleocaps) can be used to detect expression levels of viral nucleocapsid proteins and can be used to optimize detection methods for viral nucleocapsid protein levels.
Disclosure of Invention
Based on this, the object of the present invention comprises providing a specific antibody binding to coronaviruses SARS-CoV-2 and SARS-CoV nucleocapsid protein (nucleocap sideprotein) which is capable of specifically binding to nucleocapsid proteins of SARS-CoV-2 and SARS-CoV for the development of a method for detecting SARS-CoV and SARS-CoV-2 infection.
The aim of the invention can be achieved by the following technical scheme:
a first object of the present invention is to provide a specific antibody that binds to coronaviruses SARS-CoV-2 and SARS-CoV nucleocapsid protein (nucleocap sideprotein), the heavy chain CDR1 of said antibody comprising the amino acid sequence shown as SEQ ID NO.1, the heavy chain CDR2 comprising the amino acid sequence shown as SEQ ID NO.2, the heavy chain CDR3 comprising the amino acid sequence shown as SEQ ID NO. 3;
the light chain CDR1 of the antibody comprises an amino acid sequence shown as SEQ ID NO.4, the light chain CDR2 comprises an amino acid sequence GAS and the light chain CDR3 comprises an amino acid sequence shown as SEQ ID NO. 6.
Preferably, the heavy chain variable region of the antibody is shown in SEQ ID NO. 7.
Preferably, the light chain variable region of the antibody is shown in SEQ ID NO. 8.
Preferably, the light chain constant region of the antibody is shown in SEQ ID NO. 9.
Preferably, the heavy chain constant region of the antibody is shown in SEQ ID NO. 10.
Preferably, the sequence of the constant region of the antibody is the sequence of an IgG1 constant region.
Preferably, the species source of the constant region of the antibody is human.
The heavy chain variable region shown in SEQ ID No.7 and the light chain variable region shown in SEQ ID No.8 further comprise a Framework Region (FR), and the amino acid sequences of 4 FRs do not directly participate in the binding reaction.
The antibodies provided by the invention are capable of specifically binding to SARS-CoV-2 and SARS-CoV nucleocapsid proteins (nucleocaps) that are highly conserved in SARS-CoV-2 and SARS-CoV.
The second object of the present invention is to provide the application of the specific antibody combined with coronavirus SARS-CoV-2 and SARS-CoV nucleocapsid protein in SARS-CoV or SARS-CoV-2 detection or preparing medicine for resisting SARS-CoV or SARS-CoV-2.
The third object of the present invention is to provide a detection reagent, a detection kit or a medicament, comprising the above specific antibody which binds to coronavirus SARS-CoV-2 and SARS-CoV nucleocapsid protein (nucleocap apsi dprotein).
The fourth object of the present invention is to provide a nucleic acid comprising a nucleic acid sequence encoding the above antibody specific for SARS-CoV-2 and SARS-CoV nucleocapsid protein (nucleocap nucleic acid).
A fifth object of the present invention is to provide a recombinant expression vector comprising the above-mentioned nucleic acid.
Preferably, the recombinant expression vector is an antibody expression vector.
Preferably, the recombinant expression vector is AbVec2.0-IGHG1 or AbVec1.1-IGKC.
The sixth object of the present invention is to provide a host cell comprising the above specific antibody, or the above nucleic acid, or the above recombinant expression vector.
Preferably, the host cell is a 293T cell or an Expi293F cell.
The seventh object of the present invention is to provide a method for producing the above-mentioned antibody which binds to coronavirus SARS-CoV-2 and SARS-CoV nucleocapsid protein (nucleocap-si-dproteins), comprising constructing the above-mentioned host cell, culturing, and collecting the antibody which binds to coronavirus SARS-CoV-2 and SARS-CoV nucleocapsid protein (nucleocap-si-dproteins).
Preferably, the host cell is a 293T cell or an Expi293F cell.
Compared with the prior art, the technical scheme provided by the invention has at least the following beneficial effects:
the antibody provided by the invention comprises CDRs with specific sequences, can be specifically combined with coronavirus SARS-CoV-2 and SARS-CoV nucleocapsid protein, and is used for developing a SARS-CoV and SARS-CoV-2 infection detection method. The antibody provided by the invention has strong targeting and high binding activity, can specifically bind nucleocapsid proteins of SARS-CoV-2 and SARS-CoV, and has weak binding reaction with nucleocapsid proteins of another highly pathogenic coronavirus MERS-CoV. Overall, the antibodies of the invention can be used in the development of methods for the detection of SARS-CoV and SARS-CoV-2 infection. The antibody prepared by the preparation method provided by the invention has high purity and can be prepared in a large amount.
Drawings
In order to more clearly illustrate the technical solutions in the embodiments of the present application and to more fully understand the present application and its advantageous effects, the following brief description will be given with reference to the accompanying drawings, which are required to be used in the description of the embodiments. It is obvious that the drawings in the following description are only some embodiments of the present application, and that other drawings may be obtained from these drawings without inventive effort to a person skilled in the art.
FIG. 1 is a graph showing the results of detection of the binding activity of an antibody of example 2 of the present invention to SARS-CoV-2 nucleocapsid protein (Nucleocappaphprotien).
FIG. 2 is a graph showing the results of detection of the binding activity of the antibody of example 2 of the present invention to SARS-CoV nucleocapsid protein (Nucleocappside).
FIG. 3 is a graph showing the results of detection of binding activity of antibodies of example 2 of the present invention to MERS-CoV nucleocapsid protein (Nucleocappside).
Detailed Description
The present invention will be described in further detail with reference to the drawings, embodiments and examples. It should be understood that these embodiments and examples are provided solely for the purpose of illustrating the invention and are not intended to limit the scope of the invention in order that the present disclosure may be more thorough and complete. It will also be appreciated that the present invention may be embodied in many different forms and is not limited to the embodiments and examples described herein, but may be modified or altered by persons skilled in the art without departing from the spirit of the invention, and equivalents thereof are also intended to fall within the scope of the invention. Furthermore, in the following description, numerous specific details are set forth in order to provide a more thorough understanding of the invention, it being understood that the invention may be practiced without one or more of these details.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used herein in the description of the invention is for the purpose of describing the embodiments and examples only and is not intended to be limiting of the invention.
Terminology
Unless otherwise indicated or contradicted, terms or phrases used herein have the following meanings:
the term "and/or," "and/or," as used herein, includes any one of two or more of the listed items in relation to each other, as well as any and all combinations of the listed items in relation to each other, including any two of the listed items in relation to each other, any more of the listed items in relation to each other, or all combinations of the listed items in relation to each other. It should be noted that, when at least three items are connected by a combination of at least two conjunctions selected from "and/or", "or/and", "and/or", it should be understood that, in this application, the technical solutions certainly include technical solutions that all use "logical and" connection, and also certainly include technical solutions that all use "logical or" connection. For example, "a and/or B" includes three parallel schemes A, B and a+b. For another example, the technical schemes of "a, and/or B, and/or C, and/or D" include any one of A, B, C, D (i.e., the technical scheme of "logical or" connection), and also include any and all combinations of A, B, C, D, i.e., any two or three of A, B, C, D, and also include four combinations of A, B, C, D (i.e., the technical scheme of "logical and" connection).
The term "plural", and the like in the present invention refers to, unless otherwise specified, a number of 2 or more. For example, "one or more" means one kind or two or more kinds.
As used herein, "a combination thereof," "any combination thereof," and the like include all suitable combinations of any two or more of the listed items.
The "suitable" in the "suitable combination manner", "suitable manner", "any suitable manner" and the like herein refers to the fact that the technical scheme of the present invention can be implemented, the technical problem of the present invention is solved, and the technical effect expected by the present invention is achieved.
Herein, "preferred", "better", "preferred" are merely to describe better embodiments or examples, and it should be understood that they do not limit the scope of the invention. If there are multiple "preferences" in a solution, if there is no particular description and there is no conflict or constraint, then each "preference" is independent of the others.
In the present invention, "further", "still further", "particularly" and the like are used for descriptive purposes to indicate differences in content but should not be construed as limiting the scope of the invention.
In the present invention, "optional" means optional or not, that is, means any one selected from two parallel schemes of "with" or "without". If multiple "alternatives" occur in a technical solution, if no particular description exists and there is no contradiction or mutual constraint, then each "alternative" is independent.
In the present invention, the terms "first", "second", "third", "fourth", etc. are used for descriptive purposes only and are not to be construed as indicating or implying a relative importance or quantity, nor as implying an importance or quantity of a technical feature being indicated. Moreover, the terms "first," "second," "third," "fourth," and the like are used for non-exhaustive list description purposes only, and are not to be construed as limiting the number of closed forms.
In the invention, the technical characteristics described in an open mode comprise a closed technical scheme composed of the listed characteristics and also comprise an open technical scheme comprising the listed characteristics.
All documents mentioned in this application are incorporated by reference as if each were individually incorporated by reference. Unless otherwise contradicted by purpose and/or technical solution of the present application, the cited documents related to the present invention are incorporated by reference in their entirety for all purposes. When reference is made to a cited document in the present invention, the definitions of the relevant technical features, terms, nouns, phrases, etc. in the cited document are also incorporated. In the case of the cited documents, examples and preferred modes of the cited relevant technical features are incorporated into the present application by reference, but are not limited to the embodiments that can be implemented. It should be understood that when a reference is made to the description herein, it is intended to control or adapt the present application in light of the description herein.
As used herein, the term "antibody" or "immunoglobulin" is an iso-tetralin protein of about 150000 daltons, consisting of two identical light chains (L) and two identical heavy chains (H), having identical structural features. Each light chain is linked to the heavy chain by a covalent disulfide bond, while the number of disulfide bonds varies between heavy chains of different immunoglobulin isotypes. Each heavy and light chain also has regularly spaced intrachain disulfide bonds. Each heavy chain has a variable region (VH) at one end followed by a plurality of constant regions. One end of each light chain is provided with a variable region (VL) and the other end is provided with a constant region; the constant region of the light chain is opposite the first constant region of the heavy chain and the variable region of the light chain is opposite the variable region of the heavy chain. Specific amino acid residues form an interface between the variable regions of the light and heavy chains.
As used herein, the term "variable" means that certain portions of the variable regions in an antibody differ in sequence, which results in the binding and specificity of each particular antibody for its particular antigen. However, the variability is not evenly distributed throughout the antibody variable region. It is concentrated in three fragments in the light and heavy chain variable regions called Complementarity Determining Regions (CDRs) or hypervariable regions. The more conserved parts of the variable region are called Framework Regions (FR). The variable regions of the natural heavy and light chains each comprise four FR regions, which are generally in a β -sheet configuration, connected by three CDRs forming the connecting loops, which in some cases may form part of the β -sheet structure. The CDRs in each chain are held closely together by the FR regions and together with the CDRs of the other chain form the antigen binding site of the antibody [ see Kabat et al, NIH publication No.91-3242, vol. I, pp. 647-669 (1991) ]. The constant regions are not directly involved in binding of the antibody to the antigen, but they exhibit different effector functions, such as participation in antibody-dependent cytotoxicity of the antibody.
The "light chain" of a vertebrate antibody (immunoglobulin) can be classified into one of two distinct classes (called kappa and lambda) depending on the amino acid sequence of its constant region. Immunoglobulins can be classified into different classes according to the amino acid sequence of their heavy chain constant region, mainly 5 classes of immunoglobulins: igA, igD, igE, igG and IgM, some of which can be further divided into subclasses (isotypes) such as IgG1, igG2, igG3, igG4, igA and IgA2. The heavy chain constant regions corresponding to different classes of immunoglobulins are called α, δ, ε, γ, and μ, respectively. Subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known to those skilled in the art.
In general, the antigen binding properties of antibodies can be described by 3 specific regions located in the heavy and light chain variable regions, called variable regions (CDRs), which are separated into 4 Framework Regions (FRs), the amino acid sequences of the 4 FRs being relatively conserved and not directly involved in the binding reaction. These CDRs form a loop structure, the β -sheets formed by the FR therebetween are spatially close to each other, and the CDRs on the heavy chain and the CDRs on the corresponding light chain constitute the antigen binding site of the antibody. It is possible to determine which amino acids constitute the FR or CDR regions by comparing the amino acid sequences of the same type of antibody.
The invention includes not only whole antibodies but also fragments of antibodies having immunological activity or fusion proteins of antibodies with other sequences. Thus, the invention also includes fragments, derivatives and analogues of said antibodies.
In the present invention, antibodies include murine, chimeric, humanized or fully human antibodies prepared by techniques well known to those skilled in the art. Recombinant antibodies, such as chimeric and humanized monoclonal antibodies, including human and non-human portions, can be obtained by standard DNA recombination techniques, all of which are useful antibodies. Chimeric antibodies are a molecule in which different portions are derived from different animal species, e.g., chimeric antibodies having variable regions from murine monoclonal antibodies, and constant regions from human immunoglobulins (see, e.g., U.S. patent 481757 and U.S. patent 4816397, which are incorporated herein by reference in their entirety). Humanized antibodies refer to antibody molecules derived from non-human species having one or more Complementarity Determining Regions (CDRs) derived from a non-human species and a framework region derived from a human immunoglobulin molecule (see U.S. patent 5585089, incorporated herein by reference in its entirety). These chimeric and humanized monoclonal antibodies can be prepared using DNA recombination techniques well known in the art.
In the present invention, antibodies may be monospecific, bispecific, trispecific, or more multispecific.
In the present invention, the antibodies of the invention also include conservative variants thereof, meaning that up to 10, preferably up to 8, more preferably up to 5, and most preferably up to 3 amino acids are replaced by amino acids of similar or similar nature to the amino acid sequence of the antibodies of the invention to form a polypeptide. These conservatively variant polypeptides are preferably generated by amino acid substitutions according to Table A.
The following are some specific examples.
Embodiments of the present invention will be described in detail below with reference to examples. It is to be understood that these examples are illustrative of the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples, in which specific conditions are not noted, are preferably referred to the guidelines given in the present invention, and may be according to the experimental manual or conventional conditions in the art, the conditions suggested by the manufacturer, or the experimental methods known in the art.
In the specific examples described below, the measurement parameters relating to the raw material components, unless otherwise specified, may have fine deviations within the accuracy of weighing. Temperature and time parameters are involved, allowing acceptable deviations from instrument testing accuracy or operational accuracy.
In some embodiments of the invention, the host cell is a 293T cell or an Expi293F cell.
Example 1: construction, expression and purification of expression vectors for monoclonal antibodies
The monoclonal antibody prepared in this example has a heavy chain variable region comprising a heavy chain CDR1 as shown in SEQ ID NO.1, a heavy chain CDR2 as shown in SEQ ID NO.2 and a heavy chain CDR3 as shown in SEQ ID NO.3, and a light chain variable region comprising a light chain CDR1 as shown in SEQ ID NO.4, a GAS light chain CDR2 comprising the amino acid sequence and a light chain CDR3 as shown in SEQ ID NO. 6. Specifically, the heavy chain variable region is shown as SEQ ID NO.7, the light chain variable region is shown as SEQ ID NO.8, the light chain constant region is shown as SEQ ID NO.9, and the heavy chain constant region is shown as SEQ ID NO. 10.
The preparation method of the monoclonal antibody comprises the following steps:
1. referring to the published invention patent of the inventor, the patent number is: 20190975849.3 screening positive cell clones binding to coronavirus SARS-CoV-2 and SARS-CoV nucleocapsid proteins, RNA extraction of positive cell clones, using references Efficient generation ofmonoclonal antibodies from single human B cells by single cell RT-PCR and expression vector cloning. Tiller T, meffre E, yurasov S, tsuiji M, nussenzweig MC, wardemann H.J immunomethods.2008 Jan 1;329 (1-2) the heavy and light chains of the antibody were PCR amplified by the method described in 112-24.Epub 2007Oct 31.10.1016/j.jim.2007.09.017PubMed 17996249, respectively, to obtain the variable region sequences of the heavy and light chains of the antibody.
2. Nucleotide sequences encoding heavy (SEQ ID NO. 7) and light (SEQ ID NO. 8) chain variable regions of the antibody are respectively integrated into AbVec2.0-IGHG1 and AbVec1.1-IGKC (vectors are purchased from adedge functional networks, accession numbers: #80795 and # 80796), which contain the heavy and light chain constant region sequences of the human IgG1 antibody, wherein the cleavage cloning sites for AbVec2.0-IGHG1 are AgeI and SalI, and the cleavage cloning sites for AbVec1.1-IGKC are AgeI and BsiWI, respectively, and recombinant expression vectors capable of expressing the heavy and light chains of the target antibody are obtained by cleavage ligation.
3. Transfection of cells, monoclonal antibody expression and purification
1. Transfection
Transfection was performed using the Gibco Expi293F expression system (cat. No. A14635) and following the instructions, the procedure is outlined as follows:
a total of 30. Mu.g (15. Mu.g each for heavy and light chains) of two recombinant expression vector DNAs for expressing the heavy and light chains of the antibody, respectively, were used together with 80. Mu.L of the transfection reagent Expiectamine TM Mixing, and standing at room temperature for 20 minutes to form a stable compound;
then added to 25.5mL of the mixture to adjust the concentration to 2.9X10 6 cells/mL of an Expi293F cell culture medium;
at 37℃8% (v/v) CO 2 Culturing in a shaking table at 125rpm for 20 hours;
transfection enhancer 1 (150 μl) and transfection enhancer 2 (1.5 mL) carried by the Expi293F expression system were added;
continuing to place at 37 ℃ and 8% (v/v) CO 2 Culturing in a shaking table at 125rpm for 4 days.
2. Purification
The supernatant was collected by centrifugation at 3000rpm for 15 minutes, and antibody purification was performed using ProteinA magnetic beads from Genscript.
The purification steps are briefly described as follows:
500. Mu.LProteinA magnetic beads were mixed with 30mL of cell supernatant and incubated on a shaker at room temperature for 4 hours at 210RPM;
the magnetic rack adsorbs the magnetic beads, the cell supernatant is discarded, and the magnetic beads are washed 5 times with 10mL of 1 XPBS (pH 7.00.1% (v/v) Tween 20;
eluting with an Elutrionbuffer of 2.5mLpH2.00.1M glycine;
equilibrated with pH8.51M Tris buffer to pH7.0;
desalting the equilibrated monoclonal antibody with PD-10 Demulti Column (Cytiva company, cat. No. 17085101) and performing DPBS solvent replacement, eluting salt Column with 25mL of DPBS, eluting with 2.5mL of equilibrated antibody solution, and eluting with 3.5mL of DPBS to obtain monoclonal antibody solution with solvent replaced with DPBS.
The purified antibody is stored in a refrigerator at-80 ℃.
Thus, a monoclonal antibody is obtained, the heavy chain variable region of which is shown as SEQ ID NO.7, the light chain variable region is shown as SEQ ID NO.8, the light chain constant region is shown as SEQ ID NO.9, and the heavy chain constant region is shown as SEQ ID NO. 10.
Example 2: functional analysis of monoclonal antibodies
1. Detection of antigen-binding Activity of specific antibodies binding to SARS-CoV-2 and SARS-CoV nucleocapsid protein (Nucleocappaphprotein)
The binding capacity of the monoclonal antibodies obtained in example 1 to SARS-CoV-2, SARS-CoV and MERS-CoV nucleocapsid proteins (nucleocaps) was determined by ELISA.
The steps are briefly described as follows:
(1) Respectively coating 25ng of each hole of SARS-CoV-2 nucleocapsid protein (Genscript, T80101), SARS-CoV nucleocapsid protein (amino acid sequence shown as SEQ ID NO. 11) and MERS-CoV nucleocapsid protein (amino acid sequence shown as SEQ ID NO. 5) on ELISA plate, and taking DPBS (Du's phosphate buffer) as coating liquid, wherein the coating liquid is overnight at 4 ℃;
(2) DPBS of 10% (v/v) calf serum was used as a blocking solution and blocked at 37℃for 2 hours; after serial gradient dilution (10) -4 、10 -2 、10 0 、10 2 ) The monoclonal antibody to be detected prepared in example 1 of (2) was incubated at 37℃for 2 hours;
(3) HRP-conjugated Goat anti-human IgG (H+L) antibody (Jackson ImmunoResearch) diluted 1:40000 was added as secondary antibody and incubated for 1 hour at 37 ℃;
(4) After color development with TMB single-component color development solution, the reaction was terminated with 2M sulfuric acid, and the absorbance A450 value was detected with a microplate reader.
Results:
the detection result of the binding activity of the monoclonal antibody and the antigen is shown in figure 1, and the binding activity of the monoclonal antibody against SARS-CoV-2 nucleocapsid protein is shown in figure 1 as EC 50=0.0062 μg/mL; binding activity against SARS-CoV nucleocapsid protein was ec50=0.0095 μg/mL (fig. 2); binding activity against MERS-CoV nucleocapsid protein was EC50 = 1.458 μg/mL (figure 3).
The technical features of the above-described embodiments and examples may be combined in any suitable manner, and for brevity of description, all of the possible combinations of the technical features of the above-described embodiments and examples are not described, however, as long as there is no contradiction between the combinations of the technical features, they should be considered to be within the scope described in the present specification.
The above examples merely represent a few embodiments of the present invention, which facilitate a specific and detailed understanding of the technical solutions of the present invention, but are not to be construed as limiting the scope of the invention. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention. Further, it is understood that various changes and modifications of the present invention may be made by those skilled in the art after reading the above teachings, and equivalents thereof fall within the scope of the present application. It should also be understood that, based on the technical solutions provided by the present invention, those skilled in the art obtain technical solutions through logical analysis, reasoning or limited experiments, all of which are within the scope of protection of the appended claims. The scope of the patent is therefore intended to be covered by the appended claims, and the description and drawings may be interpreted as illustrative of the contents of the claims.
SEQ ID NO.1
GFTFSNAW
SEQ ID NO.2
IKSKTDGGTT
SEQ ID NO.3
TTEGSGIVVIVYAFDY
SEQ ID NO.4
QSVSSSY
SEQ ID NO.5
MASPAAPRAVSFADNNDITNTNLSRGRGRNPKPRAAPNNTVSWYTGLTQHGKVPLTFPPGQGVPLNANSTPAQNAGYWRRQDRKINTGNGIKQLAPRWYFYYTGTGPEAALPFRAVKDGIVWVHEDGATDAPSTFGTRNPNNDSAIVTQFAPGTKLPKNFHIEGTGGNSQSSSRASSLSRNSSRSSSQGSRSGNSTRGTSPGPSGIGAVGGDLLYLDLLNRLQALESGKVKQSQPKVITKKDAAAAKNKMRHKRTSTKSFNMVQAFGLRGPGDLQGNFGDLQLNKLGTEDPRWPQIAELAPTASAFMGMSQFKLTHQNNDDHGNPVYFLRYSGAIKLDPKNPNYNKWLELLEQNIDAYKTFPKKEKKQKAPKEESTDQMSEPPKEQRVQGSITQRTRTRPSVQPGPMIDVNTDSEQ ID NO.6
QQYGSSPPTSEQ ID NO.7
QVQLVQSGGGLVKPGGSLRLSCAASGFTFSNAWMSWVRQAPGKGLEWVGRIKSKTDGGTTDYAAPVKGRFTISRDDSKNTLYLQMNSLKTEDTAVYYCTTEGSGIVVIVYAFDYWGQGTLVTVSSSEQ ID NO.8
EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASRRATGIPERFSGSGSGTDFTLTINRLEPEDFAVYYCQQYGSSPPTFGGGTKVEIKSEQ ID NO.9
TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSEQ ID NO.10
STKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKSEQ ID NO.11
MSDNGPQSNQRSAPRITFGGPTDSTDNNQNGGRNGARPKQRRPQGLPNNTASWFTALTQHGKEELRFPRGQGVPINTNSGPDDQIGYYRRATRRVRGGDGKMKELSPRWYFYYLGTGPEASLPYGANKEGIVWVATEGALNTPKDHIGTRNPNNNAATVLQLPQGTTLPKGFYAEGSRGGSQASSRSSSRSRGNSRNSTPGSSRGNSPARMASGGGETALALLLLDRLNQLESKVSGKGQQQQGQTVTKKSAAEASKKPRQKRTATKQYNVTQAFGRRGPEQTQGNFGDQDLIRQGTDYKHWPQIAQFAPSASAFFGMSRIGMEVTPSGTWLTYHGAIKLDDKDPQFKDNVILLNKHIDAYKTFPPTEPKKDKKKKTDEAQPLPQRQKKQPTVTLLPAADMDDFSRQLQNSMSGASADSTQA
Claims (10)
1. A specific antibody that binds to coronavirus SARS-CoV-2 and SARS-CoV nucleocapsid proteins, comprising a heavy chain and a light chain;
the amino acid sequence of the heavy chain CDR1 of the antibody is shown as SEQ ID NO.1, the amino acid sequence of the heavy chain CDR2 is shown as SEQ ID NO.2, and the amino acid sequence of the heavy chain CDR3 is shown as SEQ ID NO. 3;
the amino acid sequence of the light chain CDR1 of the antibody is shown as SEQ ID NO.4, the amino acid sequence of the light chain CDR2 is GAS, and the amino acid sequence of the light chain CDR3 is shown as SEQ ID NO. 6.
2. The specific antibody for binding coronavirus SARS-CoV-2 and SARS-CoV nucleocapsid protein according to claim 1, wherein the heavy chain variable region of said antibody is shown in SEQ ID No. 7; and/or, the light chain variable region of the antibody is shown as SEQ ID NO. 8.
3. The specific antibody that binds to coronavirus SARS-CoV-2 and SARS-CoV nucleocapsid proteins according to claim 1, wherein the sequence of the constant region of said antibody is the sequence of the IgG1 constant region; or/and, the species source of the constant region of the antibody is human.
4. The specific antibody for binding coronavirus SARS-CoV-2 and SARS-CoV nucleocapsid protein according to claim 1, wherein the light chain constant region of said antibody is shown in SEQ ID No. 9; or/and the heavy chain constant region of the antibody is shown as SEQ ID NO. 10.
5. Use of the specific antibody binding to coronaviruses SARS-CoV-2 and SARS-CoV nucleocapsid protein of any one of claims 1 to 4 for the manufacture of a SARS-CoV or SARS-CoV-2 assay product or for the manufacture of an anti-SARS-CoV or SARS-CoV-2 medicament.
6. A test reagent, test kit or medicament comprising the specific antibody of any one of claims 1 to 4 that binds to coronavirus SARS-CoV-2 and SARS-CoV nucleocapsid proteins.
7. A nucleic acid comprising a nucleic acid sequence encoding the specific antibody of any one of claims 1 to 4 that binds to the coronaviruses SARS-CoV-2 and SARS-CoV nucleocapsid protein.
8. A recombinant expression vector comprising the nucleic acid of claim 7.
9. A host cell comprising the specific antibody of any one of claims 1 to 4, or the nucleic acid of claim 7, or the recombinant expression vector of claim 8.
10. A method for producing a specific antibody binding to coronavirus SARS-CoV-2 and SARS-CoV nucleocapsid protein as defined in any one of claims 1 to 4, comprising constructing the host cell as defined in claim 9, culturing, and collecting the specific antibody binding to coronavirus SARS-CoV-2 and SARS-CoV nucleocapsid protein.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112225797A (en) * | 2020-09-24 | 2021-01-15 | 杭州医学院 | Monoclonal antibody for resisting SARS-CoV-2 nucleocapsid protein and application thereof |
| CN114316033A (en) * | 2020-09-29 | 2022-04-12 | 香港理工大学深圳研究院 | Monoclonal antibody for specifically recognizing norovirus, preparation method and application |
| WO2023019174A2 (en) * | 2021-08-11 | 2023-02-16 | Apollos Diagnostics, Llc | Antibodies to sars-cov-2 |
| KR20230087219A (en) * | 2021-12-09 | 2023-06-16 | 한국화학연구원 | Monoclonal antibody for nucleocapsid protein of SARS-CoV-2 and uses thereof |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112225797A (en) * | 2020-09-24 | 2021-01-15 | 杭州医学院 | Monoclonal antibody for resisting SARS-CoV-2 nucleocapsid protein and application thereof |
| CN114316033A (en) * | 2020-09-29 | 2022-04-12 | 香港理工大学深圳研究院 | Monoclonal antibody for specifically recognizing norovirus, preparation method and application |
| WO2023019174A2 (en) * | 2021-08-11 | 2023-02-16 | Apollos Diagnostics, Llc | Antibodies to sars-cov-2 |
| KR20230087219A (en) * | 2021-12-09 | 2023-06-16 | 한국화학연구원 | Monoclonal antibody for nucleocapsid protein of SARS-CoV-2 and uses thereof |
Non-Patent Citations (2)
| Title |
|---|
| A SARS-CoV-2 antibody curbs viral nucleocapsid protein-induced complement hyperactivation;Sisi Kang等;《Nat Commun》;20210511;第12卷(第1期);第1-11页 * |
| 针对SARS-CoV-2核衣壳蛋白的单克隆抗体的制备和鉴定;徐双媛等;《中国人兽共患病学报》;20230118;第39卷(第01期);第20-27页 * |
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